title: Peptide specific expansion of CD8(+) T cells by recombinant plate bound MHC/peptide complexes.
abstract: Development of methods for efficient in vitro stimulation and expansion of peptide specific CD8(+) T cells is compelling not only with respect to adoptive T cell therapy but also regarding analysis of T cell responses and search for new immunogenic peptides. In the present study, a new approach to in vitro T cell stimulation was investigated. By use of an antigenic peptide derived from the cytomegalovirus (CMVp) we tested the stimulatory efficacy of recombinant plate bound MHC molecules (PB-MHC), being immobilized in culture plates. A single stimulation of non-adherent peripheral blood mononuclear cells (NA-PBMCs) with PB-MHC/CMVp resulted in significant expansion of CMVp specific CD8(+) T cells, which was comparable to that achieved by CMVp pulsed mature dendritic cells (DCs). By repeated exposure of NA-PBMCs to PB-MHC/CMVp more than 60% CMVp specific CD8(+) T cells, representing a 240-fold expansion, were reached after only two stimulations. Although stimulation with PB-MHC/CMVp clearly demonstrated efficient peptide specific expansion of CD8(+) T cells, there was a tendency to proliferative exhaustion of the cells after 3-4 stimulations. Thus, it will be of interest to examine the effect of new stimulatory cocktails, e.g. cytokines and co-stimulatory molecules, by use of the present rapid and easy-to-use method of expanding peptide specific T cells.
authors.affiliation: Laboratory of Experimental Immunology, Department of International Health, Immunology and Microbiology, the Faculty of Health Sciences, the University of Copenhagen, Copenhagen, Denmark. Esbenschmidt@gmail.com
authors.forename: Esben G W
authors.initials: EG
authors.lastname: Schmidt
authors.forename: Søren
authors.initials: S
authors.lastname: Buus
authors.forename: Mette
authors.initials: M
authors.lastname: Thorn
authors.forename: Anette
authors.initials: A
authors.lastname: Stryhn
authors.forename: Christian
authors.initials: C
authors.lastname: Leisner
authors.forename: Mogens H
authors.initials: MH
authors.lastname: Claesson
mesh_terms: D018414:CD8-Positive T-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D003587:Cytomegalovirus / Q000276:immunology*; D003713:Dendritic Cells / Q000187:drug effects / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006649:Histocompatibility Antigens / Q000276:immunology*; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000276:immunology; D008213:Lymphocyte Activation; D010750:Phosphoproteins / Q000276:immunology / Q000494:pharmacology*; D014763:Viral Matrix Proteins / Q000276:immunology / Q000494:pharmacology*
chemical_list: D006649:Histocompatibility Antigens; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa; D007371:Interferon-gamma